| 融合基因EGFRi-IL-24真核表达载体的构建与表达 |
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| 引用本文： | 王建玲,樊欣迎,包乐媛,杜连祥. 融合基因EGFRi-IL-24真核表达载体的构建与表达[J]. 生物医学工程学杂志, 2010, 0(2) |
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| 作者姓名： | 王建玲 樊欣迎 包乐媛 杜连祥 |
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| 作者单位： | 天津科技大学生物工程学院天津市工业微生物重点实验室;利兹大学分子和细胞生物学研究所; |
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| 基金项目： | 天津科技大学自然科学基金资助项目(20060206) |
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| 摘 要： | 人工合成表皮生长因子受体干扰序列(epithelial growth factor receptor interference,EGFRi),应用重叠延伸PCR(Overlapping PCR)技术将其与白细胞介素-24(IL-24)连接,并在其间引入一段柔软短肽(Gly4Ser3)。进行序列分析后,将融合基因EGFRi-IL-24重组到表达质粒pPIC9k中,Sal I酶切线性化重组质粒EGFRi-IL-24/pPIC9k,电击转化毕赤酵母GS115,经G418抗性和PCR筛选得到阳性重组菌株。重组菌株用甲醇进行诱导表达后,通过SDS-PAGE和MTT法分析鉴定蛋白表达产物。结果证实EGFRi-IL-24在毕赤酵母中获得了分泌性表达的活性蛋白,为进一步研究其生物学功能及临床应用奠定了基础。
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| 关 键 词： | 表皮生长因子受体干扰序列 毕赤酵母 蛋白表达 |
Construction of Eukaryotic Expression Vector of EGFRi-IL-24 Recombinant Gene |
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| Wang Jianling Fan Xinying Bao Leyuan Du Lianxiang. Construction of Eukaryotic Expression Vector of EGFRi-IL-24 Recombinant Gene[J]. Journal of biomedical engineering, 2010, 0(2) |
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| Authors: | Wang Jianling Fan Xinying Bao Leyuan Du Lianxiang |
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| Affiliation: | Wang Jianling1 Fan Xinying1 Bao Leyuan2 Du Lianxiang11(Tianjin Key Lab of Industrial Microbiology,College of Biotechnology,Tianjin University of Science , Technology,Tianjin 300457,China)2(Leeds Institute of Genetics,University of Leeds,UK) |
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| Abstract: | The epithelial growth factor receptor interference(EGFRi) was obtained by synthetic primers.Overlapping PCR was used to produce EGFRi-IL-24 fusion gene,which is linked by Gly4Ser3.After sequence analysis,EGFRi-IL-24 was cloned into expression vector pPIC9k;EGFRi-IL-24/pPIC9k was linearized with SacI,and then transformed to electroporated pastoris GS115.Subsequently,positive clone was selected by G418 and PCR,and its phenotype was determined by SDS-PAGE and MTT assay.The results demonstrated that EGFRi-IL-24... |
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| Keywords: | Epithelial growth factor receptor interference(EGFRi) Piehia postoris Protein expression |
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